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Team:Heidelberg/Templates/Del week11 AE
From 2013.igem.org
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(Created page with "==08-07-2013== ===Amplification from FS_02 to FS_03; 5.3 kb=== [[File:20130708 DelAE DelBB DelG.png|150px|thumb|PCR for amplification of del fragments DelAE (08.07), pSB4K5 (08.0...") |
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==08-07-2013== | ==08-07-2013== | ||
===Amplification from FS_02 to FS_03; 5.3 kb=== | ===Amplification from FS_02 to FS_03; 5.3 kb=== | ||
| - | [[File: | + | [[File:Heidelberg_20130708 DelAE DelBB DelG.png|150px|thumb|PCR for amplification of del fragments DelAE (08.07), pSB4K5 (08.07) and DelG (07.07); lane1=Marker, lanes2-4=DelAE (5.3kb), lanes5-10=pSB4K5 (4.2kb), lanes11-13:DelG (6.4kb); |
run at 100 V, 0.8 % gel (TAE)]] | run at 100 V, 0.8 % gel (TAE)]] | ||
:'''Reaction''' | :'''Reaction''' | ||
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! what !! µL | ! what !! µL | ||
|- | |- | ||
| - | | ''D. acidovorans'' || 1 | + | | ''D. acidovorans'' DSM-39|| 1 |
|- | |- | ||
| FS_02: (1/10) || 2.5 | | FS_02: (1/10) || 2.5 | ||
| Line 22: | Line 22: | ||
| - | [[File: | + | [[File:Heidelberg_20130708 DelAE DelBB DelG cut.png|150px|thumb|PCR for amplification of del fragments DelAE (08.07), pSB4K5 (08.07) and DelG (07.07), cut]] |
:'''Conditions''' | :'''Conditions''' | ||
{| class="wikitable" | {| class="wikitable" | ||
Revision as of 01:30, 30 September 2013
08-07-2013
Amplification from FS_02 to FS_03; 5.3 kb
- Reaction
| what | µL |
|---|---|
| D. acidovorans DSM-39 | 1 |
| FS_02: (1/10) | 2.5 |
| FS_03: (1/10) | 2.5 |
| Phusion Master Mix | 25 |
| DMSO | 2.5 |
| dd H2O | 19 |
- Conditions
| Biorad MyCycler | ||
|---|---|---|
| Cycles-PCR | temperature [°C] | Time [s] |
| 1 | 98 | 5 |
| 12 | 98 | 1 |
| 66 ↓ 0.5 | 5 | |
| 72 | 2:30 min | |
| 18 | 98 | 1 |
| 63 | 5 | |
| 72 | 2:30 min | |
| 1 | 72 | 10 min |
| 1 | 4 | inf |
Results:
- Amplification of DelAE worked
- bands were cut out and DNA purified using QIAquick Gel Extraction Kit
